scholarly journals THE USE OF BASIC POLYSACCHARIDES IN HISTOCHEMISTRY AND CYTOCHEMISTRY: I. THE DEMONSTRATION OF ACIDIC POLYSACCHARIDES IN TISSUE SECTIONS WITH FLUORESCEIN-LABELLED ASPERGILLUS POLYSACCHARIDE

1962 ◽  
Vol 10 (1) ◽  
pp. 8-13 ◽  
Author(s):  
E. EDWARD EVANS ◽  
SIDNEY P. KENT
Keyword(s):  
Aspergillus Parasiticus ◽  
Periodic Acid ◽  
Biological Cells ◽  
Polyvalent Cation ◽  
Periodic Acid Schiff ◽  
Tissue Sections ◽  
Formalin Fixed Paraffin ◽  
Acidic Polysaccharides ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

A procedure was described for demonstrating acidic polysaccharides in formalin-fixed paraffin-embedded tissue sections using fluorescein-labelled Aspergillus polysaccharide. This material is a basic polysaccharide composed of units of galactosamine and N-acetyl galactosamine and is synthesized by the mold Aspergillus parasiticus. The distribution of acidic polysaccharides shown by the fluorescein-labelled Aspergillus polysaccharide method was similar to results with the alcian blue-periodic acid Schiff sequence and to those with fluorescein labelled deacetylated chitin. Both epithelial and connective tissue mucins were demonstrated by the fluorescein-labelled basic polysaccharide. Aspergillus polysaccharide may be considered a colorless polyvalent cation that acts as a carrier for the fluorochrome in this procedure. Since it can react with many types of biological cells as well as with various substances isolated from cells, it may have other important applications in histochemistry and cytochemistry.

scholarly journals Immunomycology: rapid and specific immunocytochemical identification of fungi in formalin-fixed, paraffin-embedded material.

1993 ◽  
Vol 41 (8) ◽  
pp. 1217-1221 ◽  
Author(s):  
J A Reed ◽  
B A Hemann ◽  
J L Alexander ◽  
D J Brigati
Keyword(s):  
Complement Fixation ◽  
Fungal Infections ◽  
Periodic Acid ◽  
Cross Reactivity ◽  
Periodic Acid Schiff ◽  
Formalin Fixed Paraffin ◽  
Identification Of Fungi ◽  
Formalin Fixed Paraffin Embedded ◽  
Methenamine Silver ◽  
Formalin Fixed

We report the rapid (less than 1 hr), immunocytochemical identification of various fungi in formalin-fixed, paraffin-embedded tissues using antisera originally developed for use in immunodiffusion assays. Primary antisera directed towards fungal genera including Aspergillus, Blastomyces, Candida, Coccidioides, Cryptococcus, Histoplasma, and Sporothrix were examined. The specificity of each antiserum was evaluated by the presence or absence of crossreactivity with other morphologically similar fungi in both paraffin-embedded pure fungal cultures and tissues with culture-confirmed fungal infections. Each antiserum reacted strongly with the fungus to which it had been raised, whether examined in pure culture or infected tissues. The antisera raised against Candida, Cryptococcus, and Sporothrix did not exhibit cross-reactivity with any other fungus tested. However, the antisera raised to Aspergillus, Blastomyces, Coccidioides, and Histoplasma demonstrated significant crossreactivity with other genera of fungi, thus precluding their routine use in diagnostic immunocytochemistry. The results indicate that immunocytochemistry may provide an important adjunct to other methods, such as immunodiffusion or complement fixation assays and histochemical stains such as the Grocott methenamine silver or periodic acid-Schiff, when attempts are made to specifically identify certain fungi in formalin-fixed, paraffin-embedded tissues before mycology culture results are available.

scholarly journals THE USE OF BASIC POLYSACCHARIDES IN HISTOCHEMISTRY AND CYTOCHEMISTRY; II. THE DEMONSTRATION OF ACIDIC POLYSACCHARIDES IN TISSUE SECTIONS USING FLUORESCEIN-LABELLED DEACETYLATED CHITIN

1962 ◽  
Vol 10 (1) ◽  
pp. 14-18 ◽  
Author(s):  
SIDNEY P. KENT ◽  
E. EDWARD EVANS
Keyword(s):  
Connective Tissue ◽  
Acridine Orange ◽  
Alcian Blue ◽  
Periodic Acid ◽  
Fluorescent Staining ◽  
Periodic Acid Schiff ◽  
Tissue Sections ◽  
Acidic Polysaccharides ◽  
Staining Methods ◽  
Formalin Fixed

A method for demonstrating acidic polysaccharides in formalin fixed parafin embedded tissue sections using fluorescein labelled deacetylated chitin is described. Epithelial and connective tissue acidic polysaccharides have been studied in a number of organs. The distribution of acidic polysaccharides as seen with fluorescein-labelled deacetylated chitin is similar to results obtained with alcian blue-periodic acid Schiff and fluorescein-labelled Aspergillus polysaccharide. Previously reported fluorescent staining methods for acidic polysaccharides such as iron-hematoxylin-acridine orange and atabrine as well as fluorescein-labelled deacetylated chitin may prove to be useful adjuncts to the light microscopic methods of demonstrating mucins. Other possible uses of deacetylated chitin, a colorless macro-cation, in histochemistry and cytochemistry are noted.

scholarly journals Immunohistochemical Detection of Melanoma-associated Antigens on Formalin-fixed, Paraffin-embedded Canine Tumors

1994 ◽  
Vol 31 (4) ◽  
pp. 455-461 ◽  
Author(s):  
A. J. Berrington ◽  
K. Jimbow ◽  
D. M. Haines
Keyword(s):  
Monoclonal Antibodies ◽  
Periodic Acid ◽  
Specific Antigen ◽  
Periodic Acid Schiff ◽  
Cytoplasmic Staining ◽  
Normal Tissues ◽  
Melanocytic Tumors ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

Seven monoclonal antibodies (Moabs), recognizing melanoma-associated antigens in human tissues, were evaluated for their ability to immunohistochemically stain formalin-fixed, paraffin-embedded canine melanomas. Only 2 Moabs, designated human melanosome-specific antigen (HMSA)-1 and HMSA-5, stained routinely processed canine melanomas, staining 21/35 (60%) and 24/35 (69%), respectively. Twenty-nine of 35 (83%) melanomas tested were stained if results of the 2 Moabs were combined. Monoclonal antibody HMSA-1 also stained neoplastic cells of 10/35 (29%) tumors of nonmelanocytic origin and some neurons and salivary gland epithelial cells in normal canine tissues. However, Moab HMSA-1 staining in the nonmelanocytic tumors, consisting of small, discrete periodic acid-Schiff-positivc cytoplasmic droplets, was readily distinguishable from the diffusely granular, cytoplasmic staining of melanocytic tumors. In addition to melanomas, Moab HMSA-5 stained melanocytes and some melanin-containing tumor cells of a pigmented basal cell tumor and melanocytes in normal canine skin. Monoclonal antibodies HMSA-1 and HMSA-5 immunohistochemically identified the majority of canine melanomas, had limited and distinguishable staining in normal tissues and nonmelanocytic tumors, and therefore may be a useful adjunct for the diagnosis of canine melanoma in formalin-fixed, paraffin-embedded tissues.

scholarly journals Discrimination of Aspergillosis, Mucormycosis, Fusariosis, and Scedosporiosis in Formalin-Fixed Paraffin-Embedded Tissue Specimens by Use of Multiple Real-Time Quantitative PCR Assays

2016 ◽  
Vol 54 (11) ◽  
pp. 2798-2803 ◽  
Author(s):  
Elham Salehi ◽  
Mohammad T. Hedayati ◽  
Jan Zoll ◽  
Haleh Rafati ◽  
Maryam Ghasemi ◽  
...  
Keyword(s):  
Fusarium Oxysporum ◽  
Aspergillus Flavus ◽  
Ffpe Tissue ◽  
Content Type ◽  
Real Time Quantitative Pcr ◽  
Tissue Sections ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Tissue Specimens ◽  
Formalin Fixed

In a retrospective multicenter study, 102 formalin-fixed paraffin-embedded (FFPE) tissue specimens with histopathology results were tested. Two 4- to 5-μm FFPE tissue sections from each specimen were digested with proteinase K, followed by automated nucleic acid extraction. Multiple real-time quantitative PCR (qPCR) assays targeting the internal transcribed spacer 2 (ITS2) region of ribosomal DNA, using fluorescently labeled primers, was performed to identify clinically important genera and species of Aspergillus , Fusarium , Scedosporium , and the Mucormycetes . The molecular identification was correlated with results from histological examination. One of the main findings of our study was the high sensitivity of the automated DNA extraction method, which was estimated to be 94%. The qPCR procedure that was evaluated identified a range of fungal genera/species, including Aspergillus fumigatus , Aspergillus flavus , Aspergillus terreus , Aspergillus niger , Fusarium oxysporum , Fusarium solani , Scedosporium apiospermum , Rhizopus oryzae , Rhizopus microsporus , Mucor spp., and Syncephalastrum . Fusarium oxysporum and F. solani DNA was amplified from five specimens from patients initially diagnosed by histopathology as having aspergillosis. Aspergillus flavus , S. apiospermum , and Syncephalastrum were detected from histopathological mucormycosis samples. In addition, examination of four samples from patients suspected of having concomitant aspergillosis and mucormycosis infections resulted in the identification of two A. flavus isolates, one Mucor isolate, and only one sample having both R. oryzae and A. flavus . Our results indicate that histopathological features of molds may be easily confused in tissue sections. The qPCR assay used in this study is a reliable tool for the rapid and accurate identification of fungal pathogens to the genus and species levels directly from FFPE tissues.

scholarly journals Quantitative radioimmunohistochemical measurements of p185(erbB-2) in frozen tissue sections.

1996 ◽  
Vol 44 (11) ◽  
pp. 1251-1259 ◽  
Author(s):  
J R Reeves ◽  
J J Going ◽  
G Smith ◽  
T G Cooke ◽  
B W Ozanne ◽  
...  
Keyword(s):  
Tumor Biology ◽  
Breast Carcinomas ◽  
Tissue Sections ◽  
Biopsy Specimens ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Interstudy Variability ◽  
High Degree ◽  
The Relationship ◽  
Formalin Fixed

The relationship between expression of the c-erbB-2 proto-oncogene and the biology of breast cancer has been investigated widely, most studies using immunohistochemistry in formalin-fixed, paraffin-embedded tissues. This technique is at best semiquantitative and there is a high degree of interstudy variability because of its subjective nature and poor methodological standardization. The relationship between the levels of expression and biology can be examined thoroughly only with an accurately quantitative technique. We have developed a radioimmunohistochemical assay to measure p185(erbB-2) in tissue biopsy specimens. The method involves incubating frozen sections with 125I-labeled monoclonal antibody, microautoradiograpy, and grain counting with image analysis. Sections of cell pellets with known c-erbB-2 levels are processed with each batch of samples as internal calibration standards. We have quantified c-erbB-2 expression in 60 breast carcinomas and compared the results with conventional immunohistochemistry. Radioimmunohistochemistry measured receptor levels throughout the range of expression in breast carcinomas, whereas conventional immunohistochemistry detected the protein only in the highest expressing tumors. The quantitative, objective data produced by radioimmunohistochemistry allow a more thorough evaluation of the relationship between c-erbB-2 expression and tumor biology. This technique may have applications in other fields where quantitative data is required and relevant monoclonal antibodies are available.

Application of in situ hybridization with a novel phenytoin-labeled probe to conventional formalin-fixed, paraffin-embedded tissue sections

1995 ◽  
Vol 52 (3) ◽  
pp. 309-316 ◽  
Author(s):  
Yoshito Eizuru ◽  
Yoichi Minamishima ◽  
Tadashi Matsumoto ◽  
Toshinari Hamakado ◽  
Mikio Mizukoshi ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Tissue Sections ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Labeled Probe ◽  
Formalin Fixed
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